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Kit for detecting ultraviolet absorbent UV-326, and preparation method and application thereof

A UV-326, UV absorber technology, applied in measurement devices, color/spectral property measurement, instruments, etc., can solve the problems of expensive laboratory equipment, high detection cost, complicated processing, etc. Low cost, simple pre-processing effect

Active Publication Date: 2018-11-09
GUANGZHOU QUALITY SUPERVISION & TESTING INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the determination methods of ultraviolet absorbers mainly include liquid chromatography, gas chromatography-mass spectrometry, liquid chromatography-tandem mass spectrometry, etc.; pretreatment methods mainly include solid phase extraction (SPE), Soxhlet extraction (SE), rapid Solvent extraction (ASE), microwave-assisted extraction (MAE), supercritical fluid extraction (SFE), ultrasonic extraction (UE), etc.; chromatography and chromatography-mass spectrometry have the advantages of high specificity and accurate results, but these two methods The disadvantage is that the sample pretreatment is cumbersome, consumes a large amount of solvent, and requires expensive laboratory equipment, and the detection cost is high

Method used

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  • Kit for detecting ultraviolet absorbent UV-326, and preparation method and application thereof
  • Kit for detecting ultraviolet absorbent UV-326, and preparation method and application thereof
  • Kit for detecting ultraviolet absorbent UV-326, and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0043] A UV absorber UV-326 detection kit, the main components are as follows:

[0044] 1) Ultraviolet absorber UV-326 specific antigen working solution;

[0045] Prepared by the following method: 0.32g (ie 1.0mmol) of ultraviolet absorber UV-326 with phenolic hydroxyl; 0.075g (ie 1.0mmol) of 3-amino-1-propanol and 0.29g (ie 1.1mmol) of Dissolve triphenylphosphine in 2.0mL of dry tetrahydrofuran (THF), sonicate at room temperature for 5min, add 0.22mL (ie 1.1mmol) diisopropyl azodicarboxylate dropwise during sonication, continue ultrasonic reaction for 20min, after the reaction is completed Use a chromatographic column to separate the hapten of the UV absorber UV-326 that can be coupled to the protein; then mix it with N-hydroxysuccinamide (NHS) and carbodiimide (EDC), and mix it with bovine serum white Proteins were mixed, stirred at room temperature for 1 h, coupled with UV-326 specific antigen prepared by UV absorber, and diluted with concentrated sample diluent (0.01M, pH...

Embodiment 2

[0068] Establishment of an indirect competition ELISA method.

[0069] Indirect competitive ELISA method was used to detect the competitive inhibition rate of UV-326 monoclonal antibody, the method is as follows:

[0070] 1) Coat a 96-well ELISA plate with 1.5 μg / mL UV-326 specific antigen working solution, 100 μL / well, place in a 4°C refrigerator overnight, and use 250 μL / well blocking solution (5% Skimmed milk powder) was washed with washing liquid after sealing, and patted dry;

[0071] 2) Add UV absorber UV-326 standard solution (concentrations are: 2×10 5 μg / L, 2×10 4 μg / L, 2×10 3 μg / L, 2×10 2 μg / L, 2×10 1 μg / L, 2×10 0 μg / L) or 50 μL of the sample solution to be tested, then add 100 μL of UV-326 specific antibody working solution with a concentration of 3 μg / mL, mix well, and incubate at 37°C for 1 hour;

[0072] 3) After washing and patting dry, add horseradish peroxidase-labeled goat anti-mouse secondary antibody working solution diluted with diluent at a ratio o...

Embodiment 3

[0082] Kit sensitivity, specificity, accuracy.

[0083] 1. Sensitivity determination.

[0084] A standard working curve for the detection of the ultraviolet absorber UV-326 was established using the results of the indirect competitive ELISA method in Example 2. UV absorber UV-326 monoclonal antibody at 2μg / L~2×10 5 Good linearity in the range of μg / L, IC 50 =958.4ng / mL, the lowest detection limit is 5.81ng / mL, and the detection range (between 20% and 80% inhibition) is 49.50ng / mL~1650μg / mL. The detection limit of plastic food packaging products is 2.91ng / cm 2 , the detection limit of food samples is 1.16ng / g, and the detection limit of daily chemical products is 5.81ng / g. 2. Specificity Determination

[0085] Adopt the indirect competition ELISA method of embodiment 2 to measure ultraviolet absorber UV-326 structural analogue (ultraviolet absorber UV-327, ultraviolet absorber UV-234, ultraviolet absorber UV-329, ultraviolet absorber UV-71, ultraviolet absorber agent UV-3...

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Abstract

The invention relates to a kit for detecting an ultraviolet absorbent UV-326, and a preparation method and application thereof, belonging to the technical field of additive detection. The kit comprises: an ultraviolet absorbent UV-326 specific antigen working solution, wherein an ultraviolet absorbent UV-326 specific antigen is a conjugate of the ultraviolet absorbent UV-326 and a carrier protein;and an ultraviolet absorbent UV-326 specific antibody working solution, wherein an ultraviolet absorbent UV-326 specific antibody is a murine monoclonal antibody. The kit provided by the invention utilizes competitive enzyme-linked immunosorbent assay to detect the ultraviolet absorbent UV-326, and has the characteristics of high specificity, accurate results, simple pretreatment, low requirements on instruments and equipment, and low detection cost. Meanwhile, a reagent in the kit is supplied in the form of a working solution, so operation is simple and rapid; time is saved for a user; and errors caused by complicated steps are reduced. The kit provided by the invention is applicable to rapid and accurate detection of the ultraviolet absorbent UV-326 in large quantities of samples.

Description

technical field [0001] The invention relates to the technical field of additive detection, in particular to an ultraviolet absorber UV-326 detection kit and its preparation method and application. Background technique [0002] UV absorber UV-326, the chemical name is 2'-(2'-hydroxy-3'-tert-butyl-5'-methylphenyl)-5-chlorobenzotriazole, which is a kind of benzotriazole It is a representative type of ultraviolet absorber, which can effectively absorb ultraviolet rays with a wavelength of 270nm-380nm. [0003] In the field of food packaging, various packaging films, bags, barrels, boxes, bottles, cans and composite packaging materials made of plastics have been widely used, and the consumption is increasing year by year. However, under the irradiation of sunlight and some artificial light, these polymer materials are prone to auto-oxidation reaction due to the action of ultraviolet light, resulting in polymer degradation, deterioration of appearance and mechanical properties, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543G01N33/535G01N21/31
CPCG01N21/31G01N33/535G01N33/54306G01N33/577
Inventor 黄金凤刘冬虹寻知庆刘香梅郭新东汪晨霞陈立伟侯向昶吴玉銮冼燕萍
Owner GUANGZHOU QUALITY SUPERVISION & TESTING INST
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